Table IV. Comparison of FPI Parameters of 2-ABZI System with Two Other Systems Parameter
DNP (9)
2-AB21
Concentration of unbound M b 2.4 X binding sites,=R Mb Heterogeneity factor, a 0.6 0.72 Av association constant, KO 4.5 X lo7 1.4 X 1O’O Second order rate constant, 5 X 107/M 2.5 X 106/M k
Detection limit
S
10-7~
S
10-9~
Fluoresceln (70, 7 7 )
?
1 .o 1 X 10” 4 X 10*/M S
IO-I‘M
Antibody from boosted animals. In the case of fluorescein,very late antibody was used. At normal serum concentration. An inverse proportion is implied between a and the variety of binding sites. a
The F P I method can be applied to the measurement of 2ABZI with great sensitivity and specificity and indicates that this technique has a significant future potential for the analysis of environmental contaminants at the residue level. Presently, it is necessary that the contaminant be in a nearly neutral aqueous solution, and chemical processing of the material of interest may be necessary to achieve this condition. Acknowledgment Many of the detailed laboratory analyses and processes were conducted by A. Hicks of the Biophysical Chemistry Unit, Scripps Clinic and Research Foundation, La Jolla, Calif., and were carried out with considerable care and skill.
Literature Cited Discussion of Results I t is of interest to compare the FPI system for 2-ABZI to that of a typical case, as represented by dinitrophenol (DNP) (9) and its antibody, and to the optimum system evaluated to date, i.e., fluorescein and its antibody ( 1 0 , l l ) .The relevant parameters for these systems are given in Table IV. Comparison of the three systems shows that the FPI system for 2-ABZI performs better than average, but not as well as for the best test system evaluated to date, namely, fluorescein. Despite the fact that fluorescein is a considerably larger and more complex molecule than 2-ABZI, there is reason to believe that the FPI system for the latter could be brought to perform nearly as well as the fluorescein system. In particular, it seems likely that affixing carrier protein to one of the positions of the six-membered ring of 2-ABZI would provide for greater participation of the unique triple amine constellation in antibody formation relative to the present case. The present 2-ABZI-OVconjugate presents the hapten with the six-membered ring foremost in a manner that apparently minimizes the uniqueness of the molecule as it appears to the antibody-forming system. Both the small production of antibody, which suggests that less than the usual number of lymphocytes recognized the antigen as a foreign entity, and the comparatively small rate constant, k, which suggests a relatively large steric factor in the antibody-hapten reaction, are consistent with this view.
(1) Biros, F. J., Ed., “Pesticide Identification at the Residue Level”, Advances in Chemistry Series, No. 104,American Chemical Society, Washington, D.C., 1971. (2) Dandliker, W. B., “Thermodynamic and Kinetic Investigation of the Antigen-Antibody Reaction by Fluorescence Labeling Techniques”, in “Methods in Immunology and Immunochemistry”, Vol 111, L. A. Williams, and M. W. Chase, Eds., Academic Press, New York, N.Y., 1971. (3) Hawker, C. D., Anal. Chem., 45 ( l l ) ,878A (1973). (4) Centeno, E. R., Johnson, W. J., Sehon, A. H.,Int. Arch. Allergy Appl. Immunol., 3 7 , l (1970). ( 5 ) Haas, G. J., Guardia, E. J., “Production of Antibodies Against Insecticide-Protein Conjugates”, Society for Exp. Biology and Medicine, Proc., Vol 129, pp 546-51, 1968. (6) Ercegovich, C. D., “Analysis of Pesticide Residues: Immunological Techniques”, Advances in Chemistry Series, No. 104, American Chemical Society, Washington, D.C., 1971. (7) Dandliker, W. B., Kelly, R. J., Dandliker, J., Farquhar, J., Levin, J., Immunochemistry, 10,219 (1973). (8) Vaitukaitis, J., Robbins, J. B., Nieschlag, E., Ross, G. T., J . Clin. Endocrinol. Metab., 33,988 (1971). (9) Day, L. A., Sturtevant, J. M., Singer, S.J., A n n . N . Y. Acad. Sci., 103,611 (1963). (IO) Portman, A. J., Levison, S. A,, Dandliker, W. B.,Biochem. Biophys. Res. Commun., 43,207 (1971). (11) Levison, S.A., Portman, A. J., Kiertzenbaum, F., Dandliker, W. B., ibid., p 258.
Received for reuieu March 22,1976. Accepted October 7,1976. Work sponsored by the U.S. Environmental Protection Agency under Contract 68-02-1266.
NOTES
History of Heavy Metal Pollution in Southern California Coastal Zone-Reprise Kathe K. Bertine San Diego State University, San Diego, Calif. Edward D. Goldberg” Scripps Institution of Oceanography, University of California at San Diego, La Jolla, Calif. 92093 The anthropogenic fluxes of lead, vanadium, and zinc to sediments of an outer basin off the coast of Southern California, 100 km from shore, are substantially less than those to deposits 30 km from shore. The values fall off as the square of the distance from shore. Atmospheric transport, as opposed to fluvial or sewer outfall inputs, appears to be in accord with these measurements. Anthropogenic Cr, Cu, Ag, and Cd are undetectable in the outer basin.
To what distances off the coasts of highly industrialized areas are heavy metai pollutants evident in the sediments? The answer will clearly depend upon the source of the metals and their intensity as well as their modes of transport (atmosphere vs. water). In a previous study we had found that there were readily definable entries of man-mobilized lead, chromium, zinc, copper, silver, vanadium, cadmium, and m.olybdenum in sediments approximately 30 km off the Southern California coast ( I , 2). These basin deposits were anoxic, and their levels were not disturbed by the burrowing Volume
11, Number 3, March 1977
297
1 zoo
119"
118"
34O
34"
33O
33O
e
BOX CORE LOCATION
120°
1190
Figure 1. Location of box core in San Clemente Basin. In addition, flow of bottom water between basins is indicated
activities of organisms. Their rates of accumulation varied between 0.27 and 0.9 cm/year based upon Pb-210 geochronologies and varving. Anthropogenic contributions of Cr, V, and Mo in these sediments probably originated from the sewer outfall. We were unable to resolve the sources of Pb, Zn, Ag, Cd, and Cu. More recently, we have introduced a time frame into a sediment from a bank slope between San Clemente and Santa Catalina basins, 100 km to the southwest (32O51'N; 118°10'W, depth of 665 m) of the previously studied basins (Figure 1)and directed away from the centers of industrial activity ( 3 ) .The first meter of the accumulation is aerobic and appears to have been mixed by bioturbation. Its sedimentation rate is 5 cm/ 1000 years, about two orders of magnitude less than those of the inner basins. Is heavy metal pollution detectable in these deposits? Experimental The samples were analyzed by atomic absorption spectrophotometry on a Perkin-Elmer 406 unit using a deuterium background corrector where necessary. The results for Co, Ag, Cd, Pb, Cr, Cu, Mn, Ni, Zn, V, Fe, and A1 are given in Table I. The absolute concentrations of aluminum and metal/aluminum ratios are given. Aluminum is assumed to have had a uniform flux to the deposits over the past 10 000 or so years. Thus, changes in water, salt, CaC03, or organic matter contents can be compensated through normalization of the metal concentrations to that of aluminum. Discussion Of the 11 elements analyzed, three (lead, zinc, and vanadium) are significantly enriched with respect to aluminum in the uppermost 2 cm. Three other elements (cadmium, man298
Environmental Science & Technology
ganese, and cobalt) are higher in the first two levels, 0-0.5 and/or 0.5-1.0 cm, than those a t greater depth. Bioturbation resulting in downward mixing of sediments has been proposed to account for the Pu-239 240 and Pb-210 distributions ( 3 ) .For instance, Pu-239 240 is found to a depth of about 10 cm, whereas with a sedimentation rate of 5 cm/1000 years, it should be found within the top 0.2 cm or last 40 years. Anthropogenic fluxes for metals in the San Clemente deposits can be calculated by assuming that all of the higher metal values, normalized to aluminum, in the top 2 cm result from anthropogenic inputs of the metal, and by assuming that without bioturbation the higher values would have been restricted to the top 0.2 cm. This 0.2-cm level represents a time of about 1940 which is when the anthropogenic fluxes first became evident in the Inner Basin sediments. Anthropogenic fluxes for the metals in 1973 may then be calculated by determining the anthropogenic metal contents in the top 2 cm and compressing this amount into the top 0.2 cm. A linear increase is assumed from a zero value a t 0.2 cm through the calculated anthropogenic amount a t 0.1 cm to obtain the surface value. For example, the value of the Pb/Al ratio a t 0.2 cm (0) is extrapolated through the total anthropogenic PWAl ratio (1.89) a t 0.1 cm to obtain the surface value of 3.78. [The value 1.89 is the sum of the differences between the upper four PWAl ratios and the average Pb/Al ratio for the 2-22-cm interval (1.07),]Then by utilizing this value together with the sedimentation rate of 5 cm/1000 years, the average A1 content (5.46%),the solids density (assumed to be 2.6 g/cm:j), and the average solid content of 28%, the anthropogenic P b flux in 1973 of 0.04 pg/cm2/year is calculated. Table I1 compares the anthropogenic fluxes for Pb, Co, Cd, Mn, Zn, and V so derived to the San Clemente Basin with the
+
+
Table 1. Concentrations of AI and Metal/Aluminum Ratios in Units of ppm Metal/% AI in San Clemente-Santa Catalina Box Core Depth, cm
co AI
0-0.5 0.5-1.0 1.0-1.5 1.5-2.0 2.0-2.5 2.5-3.0 3.0-3.5 3.5-4.0 4.0-4.5 4.5-5.0 5-6 8-9 30-32 70-72 AVO-2
3.11 3.46 3.31 2.39 2.79 2.32 2.67 2.54 2.48 2.62 2.82 2.07 2.18 1.93 3.07f 0.48 Av2-72 2.44f 0.30 a Units of YO Fe/% AI.
Cd
Pb -
Cr -
5
AI
AI
AI
AI
AI
0.11 0.15 0.18 0.16 0.15 0.14 0.17 0.15 0.13 0.19 0.18 0.13 0.19 0.15 0.15f 0.03 0.16f 0.01
0.19 0.12 0.09 0.11 0.11 0.11 0.12 0.09 0.08 0.10 0.12 0.11 0.12 0.09 0.13f 0.04 0.11f 0.01
1.a3 1.62 1.38 1.34 1.05 1.09 1.17 1.11 0.97 1.14 1.19 1.oo 0.96 1.03 1.54% 0.23 1.07f 0.08
19.0 17.7 17.3 16.9 16.4 17.6 16.7 19.2 16.0 16.1 17.6 17.6 16.5 18.2 17.7f 0.9 17.2f 1.o
11.7 10.6 11.4 11.8 11.3 10.5 10.9 11.4 10.2 11.4 11.6
A9
10.7
10.2 9.6 11.4& 0.5 10.8f 0.7
Table II. Anthropogenic Fluxes to San Clemente Basin Compared to Anthropogenic Fluxes for Inner Basins pglcm2iyr
Element
San Clemente
Pb
0.04 0.05 0.001 0.2 0.2 0.3
co Cd Mn Zfl V a
Av Inner Barinsa
1.6
... 0.07
...
2.1 4.0
From refs. 7 and 2.
average to the Inner Basin sediments. The anthropogenic fluxes of lead, vanadium, and zinc to the San Clemente sediments are considerably less than those for the Inner Basins. Two concepts emerge from these results. First of all, the anthropogenic fluxes of zinc, vanadium, and lead from the higherly industrial Southern California coastal zone to the ocean appear to roughIy fall off as the square of the distance. The fluxes to the San Clemente Basin, 100 km from the coast, compared to those of the inner basins, about 30 km from the coast, are less by about a factor of 10 or so (Pb, 40; Zn, 10; and V, 13).This suggests that the transport of these three metals is primarily atmospheric. Surface winds prevailing off Southern California can transport atmospheric particulates in all directions. Surface ocean currents are weak and variable but sometimes are in a counterclockwise gyre around the San Clemente Basin ( 4 ) .By such a path, materials introduced to near shore surface waters off the Los Angeles area could be brought to the San Clemente region. More probably the materials would stay with the waters adjacent to the coast and move northward. The deep currents (5) between basins are
Mn -
Ni -
Zn -
AI
AI
AI
73.9 71.0 66.7 66.5 67.0 64.5 65.8 66.8 67.8 67.7 65.3 66.8 70.0 64.8 69.5f 3.7 66.6f 1.7
15.2 16.4 15.4 16.5 14.5 14.8 14.4 15.2 14.2 16.4 15.7 15.5 15.6 13.0 15.9f 0.7 14.9f 1.o
34.9 34.6 34.3 33.5 31.0 31.2 31.9 31.9 31.4 30.3 31.2 31.2 31.6 32.7 34.3f 0.6 31.4f 0.6
V Fe -
AI
21.0 17.1 14.8 15.4 9.8 11.2 14.1 11.1 12.4 15.0 15.3 14.1 12.9 10.0 17.lf 2.8 12.6% 2.0
Yo AI
AI a
0.68 0.64 0.64 0.57 0.69 0.65 0.62 0.66 0.71 0.64 0.66 0.70 0.60 0.61 0.63f 0.05 0.65f 0.04
5.47 5.49 5.44 5.44 5.74 5.61 5.62 5.52 6.06 5.35 5.67 5.81 5.50 5.71 5.46 5.66
shown in Figure 1 and, in general, move in a northwesterly direction. It is difficult to conceive of an oceanic transport to the outer basins of these highly reactive metals from the inshore zone, via the gyre, when their residence times may be of the order of months or less in coastal waters ( 1 , 2 ) . Secondly, the record of anthropogenic fluxes of heavy metals to aerobic sediments off the coast of Southern California is distorted by the burrowing actions of organisms. Pollution histories of the coastal marine environment can most readily be found in the relatively undisturbed anoxic sediments.
Acknowledgment Sonja Walawender and Wendy Goldberg assisted with some of the metal analyses. G. Michard of the University of Paris kindly provided the use of a spectrophotometer in his laboratories. Literature Cited (1) Bruland, K. W., Bertine, K., Koide, M., Goldberg, E. D., Enuiron. Sei. Technol., 8,425 (1974). (2) Bruland, K. W., Koide. M.. Goldbere. E. D.. J . GeoDhvs. Res.. 79. I
.
I
3083 (1974). (3) Koide, M., Bruland, K., Goldberg, E. D., Earth Planet. Sei. Lett., 31,31 (1976). (4) Schwartzlose, R. A., Reid, J. L., “Near-Shore Circulation in the California Current”, Calif. Mar. Res. Comm. CalCOFI Rept. 16, 56-65,1972. (5) Emery, K. O., “The Sea off Southern California”, 366 pp, Wiley, New York, N.Y., 1960.
Received for review July 6, 1976. Accepted September 13, 1976. Work supported by the Energy Research and Development Administration, Environmental Sciences Branch, Diuision of Biology and Medicine (AEC At (04-3)-34P.A. 8 4 ) .
Volume 11, Number 3, March 1977
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